Theileria mutans in Nigeria.

Two strains of bovine Theileria from northern Nigeria were shown to be identical to Theileria mutans in the indirect immunofluorescent antibody test. One of the strains was transmitted by the tick Amblyomma variegatum; large macroschizonts, typical for T mutans, could be demonstrated in infected cattle. It is concluded from these experiments and from the literature that there is reliable evidence so far for the occurrence in Nigeria of only two bovine Tehileria species, T mutans and T velifera.     

Epidemiologic aspects of scrapie and BSE including an analysis of risk factors

After a discussion of the different hypotheses about the causative agent of prion diseases, various aspects of the two most important animal prion diseases, i.e. BSE and scrapie are described. This thesis focuses on the search for a preclinical diagnosis. A major breakthrough was the discovery of a new technique for detecting the disease-associated protein in tonsillar biopsies from scrapie-infected sheep long before clinical signs appeared. Another essential part of the studies described in this thesis concerned a risk analysis for BSE in a country like the Netherlands. Major risk factors were assessed, including an assessment of the efficacy of Dutch rendering procedures in the inactivation of the agents of scrapie and BSE.     

Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

An adaptive analysis of covariance using tree-structured regression

In this article, we propose an adaptive procedure for testing for the effect of a factor of interest in the presence of one or more confounding variables in observational studies. It is especially relevant for applications where the factor of interest has a suspected causal relationship with a response. This procedure is not tied to linear modeling or normal distribution theory, and it offers a valuable alternative to traditional methods. It is suitable for applications where a factor of interest is categorical and the response is continuous. Confounding variables may be continuous or categorical. The method is comprised of two basic steps that are performed in sequence. First, confounding variables alone (i.e., without the factor of interest) are used to group observations into subsets. These subsets have the property that, when restricted to a subset, there is little or no remaining variation in the response that is attributable to the confounding variables. We then test for the factor of interest within each subset of observations. We propose to implement the first step using a technique that is generally referred to as tree-structured regression. We use a non parametric permutation procedure to carry out the second step. The proposed method is illustrated through an analysis of a U. S. Department of Agriculture (USDA) Forest Service data set and an air pollution data set.     

Ovine keratoconjunctivitis experimentally induced by instillation of Mycoplasma conjunctivae

Five sheep, free from Mycoplasma conjunctivae and ocular Chlamydia infection, were experimentally inoculated with M. conjunctivae and five more sheep were exposed to the infection by contact. Keratoconjunctivitis developed in all ten sheep. As in natural outbreaks of infectious keratoconjunctivitis (IKC), clinical signs were generally moderate and transient, and recurred in some sheep. M. conjunctivae was detected throughout the 53-day observation period. Clinical diagnosis was confirmed by histopathologic examination of three sheep. Moraxella ovis, found in six of the ten sheep before the start of the experiment, appeared to play no etiologic role in the development of IKC.     

High throughput phenotyping for aphid resistance in large plant collections

ABSTRACT: BACKGROUND: Phloem-feeding insects are among the most devastating pests worldwide. They not only cause damage by feeding from the phloem, thereby depleting the plant from photo-assimilates, but also by vectoring viruses. Until now, the main way to prevent such problems is the frequent use of insecticides. Applying resistant varieties would be a more environmental friendly and sustainable solution. For this, resistant sources need to be identified first. Up to now there were no methods suitable for high throughput phenotyping of plant germplasm to identify sources of resistance towards phloem-feeding insects. RESULTS: In this paper we present a high throughput screening system to identify plants with an increased resistance against aphids. Its versatility is demonstrated using an Arabidopsis thaliana activation tag mutant line collection. This system consists of the green peach aphid Myzus persicae (Sulzer) and the circulative virus Turnip yellows virus (TuYV). In an initial screening, with one plant representing one mutant line, 13 virus-free mutant lines were identified by ELISA. Using seeds produced from these lines, the putative candidates were re-evaluated and characterized, resulting in nine lines with increased resistance towards the aphid. CONCLUSIONS: This M. persicae-TuYV screening system is an efficient, reliable and quick procedure to identify among thousands of mutated lines those resistant to aphids. In our study, nine mutant lines with increased resistance against the aphid were selected among 5160 mutant lines in just 5 months by one person. The system can be extended to other phloem-feeding insects and circulative viruses to identify insect resistant sources from several collections, including for example genebanks and artificially prepared mutant collections.     

A serological survey of cattle in the Thames-Coromandel district of New Zealand for antibodies to Ross River virus

AIM: To determine if cattle exposed to the southern saltmarsh mosquito (SSM), Aedes camptorhynchus, in the Thames-Coromandel district of New Zealand had been exposed to Ross River virus (RRV).

METHODS: A purposive sampling design was used to test cattle from seven farms located in close proximity to four sites infested with A. camptorhynchus in the Thames-Coromandel district. Sera from 207 cattle were tested for antibodies to RRV, using an ELISA and confi rmatory virus neutralisation test (VNT) as the gold standard.

RESULTS: All 207 cattle tested negative for antibodies to RRV using the ELISA and VNT.

CONCLUSIONS: This study found no evidence of exposure to RRV in cattle in locations in the Thames-Coromandel district of New Zealand where populations of SSM were present.